Document Detail


Stable expression of the mouse nicotinic acetylcholine receptor in mouse fibroblasts. Comparison of receptors in native and transfected cells.
MedLine Citation:
PMID:  1713208     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A stable cell line expressing mouse acetylcholine receptors (AChRs), named AM4, was established by cotransfecting into NIH 3T3 fibroblasts, alpha-, beta-, gamma-, and delta-subunit cDNAs plus the neor gene by calcium phosphate precipitation. Surface AChRs on AM4 cells contain all four subunits, sediment as a single approximately 9 S peak on sucrose gradients, and have the same ratio of alpha- to beta-subunits as surface AChRs from mouse BC3H-1 cells. The surface AChRs exhibit pharmacological properties identical to those obtained for BC3H-1 cells, including the association and dissociation rates of alpha-bungarotoxin, a low affinity and cooperative instantaneous dose-response curve, cooperative steady state agonist binding and desensitization, cooperative enhancement of agonist binding affinity by local anesthetics, and distinct affinities for curariform antagonists. Patch clamp measurements on AM4 cells reveal AChR single channel properties identical to those obtained from BC3H-1 cells, including a single class of channels with a conductance of 56 pS, short and long duration openings at low and high agonist concentrations, brief and intermediate closed duration components at low agonist concentrations, and six distinct closed duration components at high agonist concentrations. The biochemical, pharmacological, and single channel measurements indicate at least 95% of the surface AChRs on AM4 cells are alpha 2 beta gamma delta pentamers.
Authors:
S M Sine; T Claudio
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Publication Detail:
Type:  In Vitro; Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  266     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  1991 Jul 
Date Detail:
Created Date:  1991-08-23     Completed Date:  1991-08-23     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  13679-89     Citation Subset:  IM    
Affiliation:
Department of Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, Connecticut 06510.
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MeSH Terms
Descriptor/Qualifier:
Animals
Blotting, Western
Bungarotoxins / metabolism
Carbachol / metabolism
Cell Line
Cloning, Molecular
Down-Regulation
Gene Expression
Ion Channel Gating
Ion Channels / physiology
Macromolecular Substances
Membrane Potentials
Mice
Molecular Weight
Parasympatholytics / metabolism
Parasympathomimetics / metabolism
Proadifen / pharmacology
Receptors, Nicotinic / chemistry,  genetics*,  physiology
Sodium / metabolism
Transfection
Grant Support
ID/Acronym/Agency:
HL38156/HL/NHLBI NIH HHS; NS21714/NS/NINDS NIH HHS
Chemical
Reg. No./Substance:
0/Bungarotoxins; 0/Ion Channels; 0/Macromolecular Substances; 0/Parasympatholytics; 0/Parasympathomimetics; 0/Receptors, Nicotinic; 302-33-0/Proadifen; 51-83-2/Carbachol; 7440-23-5/Sodium

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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